CN104730027A - Method for determining wheat stripe rust disease uredospore germination rate by utilizing near-infrared spectroscopy - Google Patents
Method for determining wheat stripe rust disease uredospore germination rate by utilizing near-infrared spectroscopy Download PDFInfo
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Abstract
The invention relates to the detection of a fungal spore germination rate and particularly provides a method for determining a wheat stripe rust disease uredospore germination rate by utilizing a near-infrared spectroscopy. According to the method, on the basis of the near-infrared spectroscopy, a judgment model of the spore germination rate is established according to the germination rate of the wheat stripe rust disease uredospore and the near-infrared spectrum data of a corresponding sample, acquired by a spore germination test method, and the automatic nondestructive detection of the wheat stripe rust disease uredospore termination rate can be realized.
Description
Technical field
The present invention relates to the detection of fungus spore germination rate, specifically, relate to a kind of method applying near-infrared spectrum technique mensuration puccinia striiformis uredospores germination rate.
Background technology
Stripe rust of wheat is the important disease of worldwide one, and this disease is wide at China's occurrence scope, repeatedly popularly causes disaster, and causes having a strong impact on to China's Wheat Production.Stripe rust of wheat relies on uredospore to carry out long-range propagation by air-flow, defines North China-northwest-middle and lower reach of Yangtze River Epidemic Flora, Yunnan Epidemic Flora and Xinjiang Epidemic Flora totally three to be very popular fauna in China.Puccinia striiformis is a kind of obligate parasite, and it, at host's endoparasitism, is constantly expanded, and shed after the uredium of generation breaks uredospore.At present, the separation of Stripe Rust, purifying, cultivation and preservation still can not be carried out on synthetic medium.For the puccinia striiformis of the work of conducting a research, be usually directly collected in the wheat leaf blade of morbidity, or come from the uredospore saved after incidence of leaf collection, sometimes also derive from the uredospore in the air collected by spore seizing device.Under field conditions (factors), owing to being subject to the impact of various environmental factor, the uredospore time-to-live is shorter; Uredospore is generally preserve under cryogenic.Uredospore only has to sprout just likely infects further and causes harm, and the great-hearted uredospore of tool has effect spread extremely important for pathogen, measures uredinial germination rate and is conducive to carrying out plant disease prevention forecast more exactly.At present, usually by (the well study of the Chinese classic of spore germination determination of test method spore germination rate, Shang Hongsheng, Li Zhenqi. Ultraviolet radiation is to the research of wheat stripe rust biological effect. Plant Pathology, 1993,23 (4): 299-304.) (Zhang Yonghong, Huang Lili, Kang Zhensheng. wheat stripe rust CY32 uredospores germination is studied. fungus journal, 2006,25 (4): 656-659.).Separately there is the report (Qiao Jiaxing of its viability of method qualitative evaluation by detecting wheat stripe rust uredospore RNA integrality, Ma Lijie, hole Sunyu, Wang Junjuan, Hu little Ping. wheat stripe rust uredospore holding time and viability evaluation. wheat crops journal, 2013,33 (5): 1043-1047.) bacterial strain that RNA integrality is good, is thought, its viability is high, otherwise the bacterial strain of RNA integrality difference, its viability is low.
But utilize spore germination determination of test method spore germination rate, waste time and energy, be subject to the impact of environment or human factor, and easily produce error when carrying out the metering of spore germination, accuracy is low.By detecting the method assessment spore viability of wheat stripe rust uredospore RNA integrality, just by the size of the integrality indirect determination uredospore viability of RNA, can not the uredinial viability of qualitative assessment.Therefore, need to seek a kind of harmless, easy, pathogen spore germination rate assay method fast and accurately.
Near-infrared spectrum technique (near infrared reflectance spectroscopy, NIRS) as a kind of quick, harmless, low cost, free of contamination analytical technology, the industries such as agricultural, food, oil, chemical industry, medicine have been widely used in.The identification of plant disease and phytopathogen can be carried out based on near-infrared spectrum technique, at present, there is no the research report carrying out pathogen spore germination rate mensuration based on near-infrared spectrum technique.
Summary of the invention
In order to solve problems of the prior art, the object of this invention is to provide a kind of method applying near-infrared spectrum technique mensuration puccinia striiformis uredospores germination rate.
In order to realize the object of the invention, technical scheme of the present invention is as follows:
Apply the method that near-infrared spectrum technique measures puccinia striiformis uredospores germination rate, it is characterized in that, it specifically comprises the steps:
1) puccinia striiformis sample is collected;
2) near infrared spectrum of puccinia striiformis sample is gathered;
3) spore germination test method is utilized to obtain the germination rate of sample;
4) near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
5) foundation of uredospores germination rate discrimination model: utilize support vector regression (supportvector regression, SVR) set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data;
6) mensuration of Stripe Rust uredospore sample germination rate to be measured: the near infrared spectrum data gathering Stripe Rust uredospore sample to be measured, after pre-service, input support vector regression discrimination model, the uredospores germination rate obtaining testing sample can be calculated.
Further, described step 3) the puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in incubator, cultivate under dark condition and sprout, then under microscopic field, each sample microscopy 200-500 uredospore, is greater than more than 1/2nd of uredospore diameter as spore germination standard using germ tube elongation, the uredinial germination rate of calculation sample.
As preferably, described step 3) the puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in 9 DEG C of incubators, cultivate under dark condition and sprout 24h, then under 20 times of microscopic fields, each sample microscopy 300 uredospores, are greater than more than 1/2nd of uredospore diameter as spore germination standard using germ tube elongation, the uredinial germination rate of calculation sample.
Further, described step 1) soak wheat seed vernalization 24h, selection is germinateed good and the artificial evenly program request of the seed that growing way is consistent is in the little basin of 10cm at diameter, every basin program request about 20, is then placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%; When wheat seedling one leaf one heart stage first, leaf launched completely, carry out the artificial spray inoculation of stripe rust of wheat; From the liquid nitrogen container preserving germ, take out required Stripe Rust biological strain, 40 DEG C of water-bath 5min, then 4 DEG C of dark aquation 12h, are made into spore suspension by the Tween-80 of proper amount of strains and 0.2%; Remove blade top layer wax with finger-dipping clear water, then carry out artificial spray inoculation, inoculation is placed on moisturizing 24h under the dark condition of 11 ~ 13 DEG C; Finally be placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%; Collect Stripe Rust uredospore after treating wheat seedling morbidity, and be placed in the exsiccator under 4 DEG C of conditions and save backup.
Further, described step 2) before collection spectrum, the uredospore preserving different time is mixed at random, to obtain germination rate between 0% ~ 100% and its equally distributed as far as possible uredospore sample, sample size is not less than 30;
During spectra collection, be divided into by each puccinia striiformis uredospore sample and be not less than 3 parts, put into measuring cup, utilize integrating sphere diffuse reflection methodology to gather the near infrared light spectrum information of puccinia striiformis, spectral range is 4000 ~ 12000cm
-1, spectral resolution is not less than 16cm
-1, scanning times is no less than 16 times, using the spectrum as this sample after the spectrum of each for same sample part is average.
Further, described step 5) Spectral range selection 8000 ~ 11000cm
-1, utilize support vector regression to set up uredospores germination rate discrimination model.
Present invention also offers a kind of puccinia striiformis uredospores germination rate discrimination model, the method for building up of described model is:
S1: collect puccinia striiformis sample;
S2: the near infrared spectrum gathering puccinia striiformis sample;
S3: utilize spore germination test method to obtain the germination rate of sample;
S4: near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
S5: the foundation of uredospores germination rate discrimination model: utilize support vector regression to set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data.
Further, the method for building up of described model is:
S1: collect puccinia striiformis sample:
When wheat seedling one leaf one heart stage first, leaf launched completely, carry out the artificial spray inoculation of stripe rust of wheat; From the liquid nitrogen container preserving germ, take out required Stripe Rust biological strain, 40 DEG C of water-bath 5min, then 4 DEG C of dark aquation 12h, are made into spore suspension by the Tween-80 of proper amount of strains and 0.2%; Remove blade top layer wax with finger-dipping clear water, then carry out artificial spray inoculation, inoculation is placed on moisturizing 24h under the dark condition of 11 ~ 13 DEG C; Finally be placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%; Collect Stripe Rust uredospore after treating wheat seedling morbidity, and be placed in the exsiccator under 4 DEG C of conditions and save backup;
S2: the near infrared spectrum gathering puccinia striiformis sample:
Before collection spectrum, mix preserving the uredospore of different time at random, to obtain germination rate between 0% ~ 100% and its equally distributed as far as possible uredospore sample, sample size is not less than 30;
During spectra collection, be divided into by each puccinia striiformis uredospore sample and be not less than 3 parts, put into measuring cup, utilize integrating sphere diffuse reflection methodology to gather the near infrared light spectrum information of puccinia striiformis, spectral range is 4000 ~ 12000cm
-1, spectral resolution is not less than 16cm
-1, scanning times is no less than 16 times, using the spectrum as this sample after the spectrum of each for same sample part is average;
S3: utilize spore germination test method to obtain the germination rate of sample:
The puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in 9 DEG C of incubators, cultivate under dark condition and sprout 24h, then under 20 times of microscopic fields, each sample microscopy 300 uredospores, more than 1/2nd of uredospore diameter are greater than as spore germination standard, the uredinial germination rate of calculation sample using germ tube elongation;
S4: near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
S5: the foundation of uredospores germination rate discrimination model:
Spectral range selects 8000 ~ 11000cm
-1, utilize support vector regression to set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data.
Present invention also offers described model and measure the application in puccinia striiformis uredospores germination rate.
Particularly, described in be applied as the near infrared spectrum data gathering Stripe Rust uredospore sample to be measured, input after pre-service in foregoing model, calculate and obtain the uredospores germination rate of testing sample.
Beneficial effect of the present invention is:
The present invention is based on near-infrared spectrum technique, the uredinial germination rate of wheat stripe rust obtained according to spore germination test method and the near infrared spectrum data of respective sample thereof, set up the discrimination model of spore germination rate, achieve the automatic Non-Destructive Testing of puccinia striiformis uredospores germination rate.
The present invention utilizes the discrimination model of foundation to measure pathogen spore germination rate, the quick nondestructive that can be used for puccinia striiformis uredospores germination rate measures, also the quick nondestructive discrimination model that the present invention sets up other pathogen spore germination rates can be applied, provide fast a kind of, harmless, low cost, pollution-free, the analytical technology that accuracy is high, the time of carrying out pathogen spore germination test in a large number can be saved, and achieve the quantitative Fast Measurement of pathogen spore germination rate, and can in the limited situation of pathogen spore amount, save pathogen spore, ensure other carrying out tested.
Accompanying drawing explanation
Fig. 1 is 64 the near infrared light spectral curves gathering acquisition in the embodiment of the present invention 1.
Fig. 2 is modeling collection (a) of institute's established model in the embodiment of the present invention 1 and the actual value of test set (b) and predicted value graph of a relation.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
1, collect puccinia striiformis sample: wheat breed is engraved virtuous 169 seed immersion vernalization 24h, selection is germinateed good and the artificial evenly program request of the seed that growing way is consistent is in the little basin of 10cm at diameter, every basin program request about 20, is then placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%.When wheat seedling one leaf one heart stage first, leaf launched completely, carry out the artificial spray inoculation of stripe rust of wheat.From the liquid nitrogen container preserving germ, take out required Stripe Rust biological strain, 40 DEG C of water-bath 5min, then 4 DEG C of dark aquation 12h, are made into spore suspension by the Tween-80 of proper amount of strains and 0.2%.Remove blade top layer wax with finger-dipping clear water, then carry out artificial spray inoculation, inoculation is placed on moisturizing 24h under the dark condition of 11 ~ 13 DEG C.Finally be placed in the indoor cultivation of the artificial climate with above-mentioned environmental parameter.Collect Stripe Rust uredospore after treating wheat seedling morbidity, and be placed in the exsiccator under 4 DEG C of conditions and save backup.Different batches need be divided to carry out numerous bacterium, receive bacterium, cause the difference of uredospores germination rate to ensure the difference of holding time.The enough research institutes of bacterium amount to be deposited take the collection of the near infrared light spectral curve carrying out pathogen.
2, near infrared spectrum data collection: before collection spectrum, mixes at random by preserving the uredospore of different time, to obtain germination rate between 0% ~ 100% and its equally distributed as far as possible uredospore sample.The wheat stripe rust uredospore sample of 64 different germination rates is obtained altogether, each sample 160mg in research.The instrument gathering puccinia striiformis near infrared spectrum Curves is the MPA Fourier near infrared spectrometer that German Bruker company produces.During spectra collection, each puccinia striiformis uredospore sample average is divided into 4 parts, every part of 40mg, first, a copy of it sample is put into the measuring cup that diameter is 4mm, keeps its tightness consistent as far as possible, to reduce the test error caused because its tightness is different.Utilize integrating sphere diffuse reflection methodology to gather the near infrared light spectrum information of puccinia striiformis, spectral range is 4000 ~ 12000cm
-1, spectral resolution is 8cm
-1, scanning times is 32 times.Then, gather all the other 3 parts of sample spectra by same method, so, each sample gathers 4 spectrum altogether, is averaging the spectrum as this sample to 4 spectrum, obtains 64 near infrared spectrums altogether, as shown in Figure 1.
3, the acquisition of sample germination rate data: each puccinia striiformis uredospore sample after gathering spectrum is mixed with 0.1% water agar respectively, and be placed in 9 DEG C of incubators, cultivate under dark condition and sprout 24h, then under 20 times of microscopic fields, each sample microscopy 300 uredospores, be greater than more than 1/2nd of uredospore diameter as spore germination standard using germ tube elongation, calculate the uredinial germination rate of each sample.After obtaining the germination rate of each sample, the curve of spectrum of collection and the spore germination rate of respective sample are incorporated into the data analysis of line correlation.
4, the pre-service of near infrared spectrum data: for selecting suitable near infrared spectrum preprocess method, bd2 small echo 1 layer is adopted to decompose denoising, bd2 small echo 2 layers decomposes denoising, bd2 small echo 3 layers decomposes denoising, normalization (normalization), additional dispersion corrects (multiplicationscatter correction, MSC), standard normal variable conversion (standard normalizedvariate, SNV), vector normalization (vector normalization, VN), single order convolution differentiate (Savitzky-Golay first derivative) and Second Order Convolution differentiate (Savitzky-Golaysecond derivative) amount to 9 kinds of methods and carry out pre-service to spectrum.Wherein, utilize Savitzky-Golay method to calculate single order and lead when leading with second order, window size selects 7, and the degree of polynomial selects 3.More than calculate and all realize in MATLAB7.8.0 (R2009a).The wavelet toolbox using MATLAB to carry carries out Wavelet Denoising Method process, and Wavelet noise-eliminating method selects soft threshold method (soft thresholding), and heuristic Threshold selection method (heursure) is selected in the determination of threshold value.Function call form and parameter as follows:
wt＝wden(x,'heursure','s','one',N,'db2')
Wherein, wt is the spectrum after Wavelet Denoising Method; X is original spectrum; N is Decomposition order.
5, the foundation of uredospores germination rate discrimination model: all samples is by being modeling collection and test set according to the ratio cut partition of 1:1,2:1,3:1,4:1 and 5:1 respectively after the sequence of germination rate gradient.For selecting the spectrum district of applicable modeling, by 4000 ~ 12000cm
-1wave band is divided into 36 Zhong Pu districts, respectively: 4000 ~ 5000, 4000 ~ 6000, 4000 ~ 7000, 4000 ~ 8000, 4000 ~ 9000, 4000 ~ 10000, 4000 ~ 11000, 4000 ~ 12000, 5000 ~ 6000, 5000 ~ 7000, 5000 ~ 8000, 5000 ~ 9000, 5000 ~ 10000, 5000 ~ 11000, 5000 ~ 12000, 6000 ~ 7000, 6000 ~ 8000, 6000 ~ 9000, 6000 ~ 10000, 6000 ~ 11000, 6000 ~ 12000, 7000 ~ 8000, 7000 ~ 9000, 7000 ~ 10000, 7000 ~ 11000, 7000 ~ 12000, 8000 ~ 9000, 8000 ~ 10000, 8000 ~ 11000, 8000 ~ 12000, 9000 ~ 10000, 9000 ~ 11000, 9000 ~ 12000, 10000 ~ 11000, 10000 ~ 12000 and 11000 ~ 12000cm
-1.Apply different preprocess methods, modeling ratio (modeling collection: test set) and spectral range and set up support vector regression (SVR) model, select to differentiate that the good model of effect is as the discrimination model detecting puccinia striiformis uredospores germination rate.Select radial basis function (radial basis function, RBF) as kernel function modeling, use grid-search algorithms (grid search algorithm) search optimum punishment parameter C and kernel functional parameter g, hunting zone is 2
-8~ 2
8, search step pitch is 0.8, and in traversal grid, there is a computation model square error (mean squared error, MSE) in institute, and Search Results when selecting square error minimum is as model parameter.Call the coefficient of determination (R
2) and the prediction level of square error evaluation model and repeatability.Model result good under different pretreatments is listed in table 1.Model checking effect listed in table 1 is all better, and in these models, comparatively speaking, preprocess method selects vector normalization, and modeling is than being 5:1, and Spectral range selects 8000 ~ 11000cm
-1time institute's established model modeling collection coefficient of determination is relatively high, and square error is relatively little, and the test set coefficient of determination is maximum, and square error is minimum, and Spectral range used is minimum, therefore thinks that this forecast result of model is best.Fig. 2 shows, this model can predict the germination rate of Stripe Rust sample well.Therefore, select this model as the discrimination model of Stripe Rust uredospores germination rate.
Best model result under table 1 different pretreatments method
Modeling collection (a) of Fig. 2 institute established model and the actual value of test set (b) and predicted value graph of a relation
6, the mensuration of Stripe Rust uredospore sample germination rate to be measured; Gather the near infrared spectrum data of Stripe Rust uredospore sample to be measured, after pre-service, input discrimination model, can calculate and obtain uredinial germination rate.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (10)
1. apply the method that near-infrared spectrum technique measures puccinia striiformis uredospores germination rate, it is characterized in that, it specifically comprises the steps:
1) puccinia striiformis sample is collected;
2) near infrared spectrum of puccinia striiformis sample is gathered;
3) spore germination test method is utilized to obtain the germination rate of sample;
4) near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
5) foundation of uredospores germination rate discrimination model: utilize support vector regression to set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data;
6) mensuration of Stripe Rust uredospore sample germination rate to be measured: the near infrared spectrum data gathering Stripe Rust uredospore sample to be measured, after pre-service, input support vector regression discrimination model, the uredospores germination rate obtaining testing sample can be calculated.
2. method according to claim 1, it is characterized in that, described step 3) the puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in incubator, cultivate under dark condition and sprout, then under microscopic field, each sample microscopy 200-500 uredospore, more than 1/2nd of uredospore diameter are greater than as spore germination standard, the uredinial germination rate of calculation sample using germ tube elongation.
3. method according to claim 2, it is characterized in that, described step 3) the puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in 9 DEG C of incubators, cultivate under dark condition and sprout 24h, then under 20 times of microscopic fields, each sample microscopy 300 uredospores, more than 1/2nd of uredospore diameter are greater than as spore germination standard, the uredinial germination rate of calculation sample using germ tube elongation.
4. the method according to any one of claim 1-3, is characterized in that, described step 1) when wheat seedling one leaf one heart stage first, leaf launched completely, carry out the artificial spray inoculation of stripe rust of wheat; Inoculation is placed on moisturizing 24h under the dark condition of 11 ~ 13 DEG C; Finally be placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%; Collect Stripe Rust uredospore after treating wheat seedling morbidity, and be placed in the exsiccator under 4 DEG C of conditions and save backup.
5. method according to claim 4, it is characterized in that, described step 2) before collection spectrum, the uredospore preserving different time is mixed at random, to obtain germination rate between 0% ~ 100% and equally distributed as far as possible uredospore sample, sample size is not less than 30;
During spectra collection, be divided into by each puccinia striiformis uredospore sample and be not less than 3 parts, put into measuring cup, utilize integrating sphere diffuse reflection methodology to gather the near infrared light spectrum information of puccinia striiformis, spectral range is 4000 ~ 12000cm
-1, spectral resolution is not less than 16cm
-1, scanning times is no less than 16 times, using the spectrum as this sample after the spectrum of each for same sample part is average.
6. method according to claim 5, is characterized in that, described step 5) Spectral range selection 8000 ~ 11000cm
-1, utilize support vector regression to set up uredospores germination rate discrimination model.
7. puccinia striiformis uredospores germination rate discrimination model, is characterized in that, the method for building up of described model is:
S1: collect puccinia striiformis sample;
S2: the near infrared spectrum gathering puccinia striiformis sample;
S3: utilize spore germination test method to obtain the germination rate of sample;
S4: near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
S5: the foundation of uredospores germination rate discrimination model: utilize support vector regression to set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data.
8. model according to claim 7, is characterized in that, the method for building up of described model is:
S1: collect puccinia striiformis sample:
When wheat seedling one leaf one heart stage first, leaf launched completely, carry out the artificial spray inoculation of stripe rust of wheat; From liquid nitrogen container, take out required Stripe Rust biological strain, 40 DEG C of water-bath 5min, then 4 DEG C of dark aquation 12h, are made into spore suspension by the Tween-80 of proper amount of strains and 0.2%; Remove blade top layer wax with finger-dipping clear water, then carry out artificial spray inoculation, inoculation is placed on moisturizing 24h under the dark condition of 11 ~ 13 DEG C; Finally be placed in the indoor cultivation of artificial climate that environmental parameter is 12h illumination, intensity of illumination 10000lux, temperature 11 ~ 13 DEG C, relative humidity 60% ~ 70%; Collect Stripe Rust uredospore after treating wheat seedling morbidity, and be placed in the exsiccator under 4 DEG C of conditions and save backup;
S2: the near infrared spectrum gathering puccinia striiformis sample:
Before collection spectrum, mix preserving the uredospore of different time at random, to obtain germination rate between 0% ~ 100% and its equally distributed as far as possible uredospore sample, sample size is not less than 30;
During spectra collection, be divided into by each puccinia striiformis uredospore sample and be not less than 3 parts, put into measuring cup, utilize integrating sphere diffuse reflection methodology to gather the near infrared light spectrum information of puccinia striiformis, spectral range is 4000 ~ 12000cm
-1, spectral resolution is not less than 16cm
-1, scanning times is no less than 16 times, using the spectrum as this sample after the spectrum of each for same sample part is average;
S3: utilize spore germination test method to obtain the germination rate of sample:
The puccinia striiformis uredospore sample having gathered near infrared spectrum is mixed with 0.1% water agar respectively, and be placed in 9 DEG C of incubators, cultivate under dark condition and sprout 24h, then under 20 times of microscopic fields, each sample microscopy 300 uredospores, more than 1/2nd of uredospore diameter are greater than as spore germination standard, the uredinial germination rate of calculation sample using germ tube elongation;
S4: near infrared spectrum data pre-service: select vector method for normalizing to carry out pre-service to spectrum;
S5: the foundation of uredospores germination rate discrimination model:
Spectral range selects 8000 ~ 11000cm
-1, utilize support vector regression to set up uredospores germination rate discrimination model, determine the quantitative relationship between uredospores germination rate and near infrared spectrum data.
9. the model described in claim 7 or 8 is measuring the application in puccinia striiformis uredospores germination rate.
10. application according to claim 9, is characterized in that, gathers the near infrared spectrum data of Stripe Rust uredospore sample to be measured, inputs the model described in claim 7 or 8 after pre-service, calculates the uredospores germination rate obtaining testing sample.
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| CN105803070A (en) * | 2016-02-05 | 2016-07-27 | 中国农业大学 | Method for measuring relative content of puccinia striiformis DNA (deoxyribose nucleic acid) in wheat leaves |
| CN105803070B (en) * | 2016-02-05 | 2019-05-03 | 中国农业大学 | Stripe Rust DNA relative amount measurement method in a kind of wheat leaf blade |
| CN108760647A (en) * | 2018-03-20 | 2018-11-06 | 南京财经大学 | A kind of wheat content of molds line detecting method based on Vis/NIR technology |
| CN109389581A (en) * | 2018-09-10 | 2019-02-26 | 江苏大学 | The method of wheat leaf rust classification based on infrared thermal imaging edge detection process |
| CN113567392A (en) * | 2021-07-20 | 2021-10-29 | 西北农林科技大学 | Wheat airborne pathogenic bacterium spore rapid nondestructive identification method based on near infrared spectrum |
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